1.Leptospira interrogans inducing apoptosis of macrophages through mitochondria-associated signaling pathway
Xingli FAN ; Haiyan DONG ; Jie YAN
Chinese Journal of Microbiology and Immunology 2009;29(9):807-812
Objective To determine mitochondria-associated signaling pathway on cell apoptosis in Leptospira interrogans infected murine mononuclear-macrophages. Methods A cell apoptotic model of L. interrogan serogroup Icterohaemorrhagiae strain Lai inducing apoptosis of murine mononuclear-macrophage line (J774A. 1) was established. Pathological changes of mitochondria in the infected cells were observed under transmission electron microscope. Mitochondrial potential and reactive oxygen species(ROS) levels in the infected cells were detected using JC-1 staining method and DCFH-DA fluorescent probe, respectively,and caspase-8/-9 activities in the infected cells were measured using commercial kits. and the apoptosis block effects of caspase inhibitors were determined by flow cytometry. Apoptosis in the infected cells and ap-optosis-blocking effect by caspase inhibitors were detected by flow cytometry. Western blot assay was adopted to examine the levels of cytochrome C (CytC) , AIF, EndoG and Smae in the mitochondria and cytoplasm.The transposition of either AIF or EndoG from the cytoplasm into cell nucleus was determined by immunoflu-orescence staining test. Results L. interrogans strain Lai could induce J774A. 1 cell apoptoais. In the in-fected cells, visible mitochondrial injury, declined mitochondrial membrane potential and elevated ROS level were presented. The activity of caspase-8 but not of caspase-9 was significantly increased. The caspase in-hihitors could not completely block the cell apoptosis. Both the AIF and EndoG was released from the mito-chondria and subsequently transferred from the cytoplasm into the nucleus in the infected cells. However,elevation of CytC level and Smac release in cytoplasm of the infected cells could not be found. Conclusion L.interrogans can induce apoptosis in the infected mononuclear-macrophage via AIF and EndoG belonging to caspase-independent pathway.
2.Efficacy of ultrasound-guided transversus abdominis plane block with different concentrations of ropivacaine for analgesia after cesarean section
Yan DONG ; Qing ZHANG ; Fan HUANG
Chinese Journal of Anesthesiology 2015;(12):1449-1451
Objective To evaluate the efficacy of ultrasound?guided transversus abdominis plane ( TAP) block with different concentrations of ropivacaine for analgesia after cesarean section. Methods A total of 120 parturients, aged 24-31 yr, of American Society of Anesthesiologists physical statusⅠ or Ⅱ, weighing 64-73 kg, at 35 to 41 week gestation, scheduled for elective cesarean section, were randomly di?vided into 3 groups ( n= 40 each) using a random number table: 0?25% ropivacaine group ( group Ⅰ) , 0?20% ropivacaine group ( groupⅡ) , and 0?15% ropivacaine group ( groupⅢ) . A patient?controlled an?algesia pump was connected at the end of surgery, and the corresponding concentration of ropivacaine 1?5 mg∕kg was injected into the bilateral TAP under the guidance of ultrasound in each group. Visual analogue scale score was maintained ≤3. The number of attempt and the number of patients requiring rescue analge?sic were recorded at 48 h after surgery. The occurrence of TAP block?related complications was observed and recorded. Results Compared with groupⅢ, the number of attempt and the number of patients requi?ring rescue analgesic were significantly decreased in Ⅰ and Ⅱ groups ( P<0?05 ) . Compared with groupⅡ, the number of attempt and the number of patients requiring rescue analgesic were significantly de?creased in group Ⅰ (P<0?05). No TAP block?related complications were detected in the three groups. Conclusion Ultrasound?guided TAP block with 0?25% ropivacaine is helpful in improving the analgesic efficacy after cesarean section without serious complications.
3.Determination of baicalin of scutellaria-extract by FT-NIRS rapidly
Yan BAI ; Le LIU ; Dong WANG ; Kefeng FAN
Chinese Traditional Patent Medicine 1992;0(09):-
AIM: To determine baicalin of scutellaria-extract rapidly by FT-NIRS and data analysis software. METHODS: The correction model was set up based on partial least square and was used to predict baicalin of scutellaria-extract in the samples. RESULTS: Cross validation and test samples determination showed that the co-rrelation coefficient(R2) of this correction model was 95.05,the RMSECV was 0.861,respectively. CONCLUSION: It is fast and convenient.The correction model could be used to predict baicalin of scutellaria-extract rapidly.It can offer reference to content determination of other extracts of Chinese herbs.
4.Time-series Analysis in Imatinib-resistant Chronic Myeloid Leukemia K562-cells under Different Drug Treatments
ZHAO YAN-HONG ; ZHANG XUE-FANG ; ZHAO YAN-QIU ; BAI FAN ; QIN FAN ; SUN JING ; DONG YING
Journal of Huazhong University of Science and Technology (Medical Sciences) 2017;37(4):621-627
Chronic myeloid leukemia (CML) is characterized by the accumulation of active BCR-ABL protein.Imatinib is the first-line treatment of CML;however,many patients are resistant to this drug.In this study,we aimed to compare the differences in expression patterns and functions of time-series genes in imatinib-resistant CML cells under different drug treatments.GSE24946 was downloaded from the GEO database,which included 17 samples of K562-r cells with (n=12) or without drug administration (n=5).Three drug treatment groups were considered for this study:arsenic trioxide (ATO),AMN107,and ATO+AMN107.Each group had one sample at each time point (3,12,24,and 48 h).Time-series genes with a ratio of standard deviation/average (coefficient of variation) >0.15 were screened,and their expression patterns were revealed based on Short Time-series Expression Miner (STEM).Then,the functional enrichment analysis of time-series genes in each group was performed using DAVID,and the genes enriched in the top ten functional categories were extracted to detect their expression patterns.Different time-series genes were identified in the three groups,and most of them were enriched in the ribosome and oxidative phosphorylation pathways.Time-series genes in the three treatment groups had different expression patterns and functions.Time-series genes in the ATO group (e.g.CCNA2 and DAB2)were significantly associated with cell adhesion,those in the AMN107 group were related to cellular carbohydrate metabolic process,while those in the ATO+AMN107 group (e.g.AP2M1) were significantly related to cell proliferation and antigen processing.In imatinib-resistant CML cells,ATO could influence genes related to cell adhesion,AMN107 might affect genes involved in cellular carbohydrate metabolism,and the combination therapy might regulate genes involved in cell proliferation.
5.Effects of chlorzoxazone on HepG2 tumor cells
Changlong FAN ; Yan LUAN ; Yan CHEN ; Jun ZHANG ; Wan ZHANG ; Baofeng YANG ; Deli DONG
Chinese Pharmacological Bulletin 2003;0(09):-
Aim To investigate effects of chlorzoxazone on survival and apoptosis of HepG2 cells.Methods The necrosis of HepG2 cells was evaluated by measurement of LDH release.The effects of chlorzoxazone on survival of HepG2 cells were assayed by MTT dyereduction.The effects of chlorzoxazone on cell apoptosis was analyzed by TUNEL method.The ultrastructure of HepG2 cells was observed by transmission electron microscope.Results Chlorzoxazone at concentrations of 100~500 ?mol?L-1 inhibited survival ratios of HepG2 cells in a dose-dependent manner significantly.Typical apoptotic changes were observed in HepG2 cells under the fluorescence microscope and transmission electron microscope.Apoptosis of HepG2 cells was induced after treatment of chlorzoxazone at concentrations from 100 ?mol?L-1 to 500 ?mol?L-1 for 48h,which showed obvious concentration-effect relationship.The apoptotic ratios of HepG2 cells were also increased when chlorzoxazone(100,200,300 and 500 ?mol?L-1) was treated for 24,48 and 72 h,which showed obvious time-effect relationship.Conclusion Chlorzoxazone inhibited HepG2 cells survival and induced cell apoptosis.
6.Resistance Phenotype and Efflux Protein Gene Expression:Study of 51 Strains of Acinetobacter Baumannii
Tao DONG ; Rui WANG ; Weihang TONG ; Dong CHAI ; Fei PEI ; Beibei LIANG ; Yan FAN ; Congran LI
China Pharmacy 2005;0(13):-
OBJECTIVE: To investigate the resistance phenotype of acinetobacter baumannii and the expression of ade efflux pump gene. METHODS: Non-repetitive 51 strains of Acinetobacter baumannii were collected in Peking Union Medical Hospital between Feb. 2004 and Feb. 2005. The active efflux system adeB structural gene and sequence were identified by PCR. RESULTS: The tested strains were not susceptible to common broad-spectrum antibiotics. Multidrug resistant Acinetobacter baumannii carried adeB active efflux pump gene. CONCLUSION: The multiple-drug resistance of Acinetobacter baumannii is independent of ?-lactamase. It maybe related to other drug resistance mechanism. The effect of active efflux system is possibly one of the major mechanisms of multidrug resistance of acinetobacter baumannii.
7.Study on apoptosis of human ovarian cancer cell induced by siRNA on Rab25
Yang FAN ; Xiao-Yan XIN ; Bi-Liang CHEN ; Xiang-Dong MA ; Li SHANG ;
Cancer Research and Clinic 1999;0(05):-
Objective To study the effect of siRNA of Rab25 on apoptosis induction in ovarian carci- noma cell.Methods According to Rab25 mRNA sequence in the Genebank,the plasmids expressing siRNA against Rab25 were constructed and stably transected into A2780 cells,MTY method were applied to measure cell growth and proliferation.Apoptosis rate and cell cycle phase distribution of A2780 cells were measured by flow cytometry(FCM).Apoptosis was confirmed by agarose gel electrophoresis(AGE)of DNA.Results Cells transected with the plasmids expressing siRNA targeting Rab25 gene effectively decreased cell growth, proliferation;blocked A2780 cells in the G_1 phase of cell cycle and induced cell apoptosis.A typical DNA ladder pattern appeared on AGE.Conclusion Rab25 gene siRNA can inhibit growth and induce apoptosis in ovarian carcinoma cell line,which will facilitate further studies of Rab25 function and its application in the treatment of ovarian cancer.
8.The effect of finofibrate and simvastatin on the serum free fatty acids of alcoholic fatty liver in rats
Ming YAN ; Fan-Li MENG ; Chun-Xia DONG ; Rui-Juan LV ; Xiao-Qing JIA ;
Chinese Pharmacological Bulletin 2003;0(11):-
AIM To investigate the effect of fenofibrate and simvastatin on the serum free fatty acids of alcoholic fatty liver in rats. METHODS The rat model of alcoholic fatty liver was reproduced by chronic ethanol ingestion plus olive oil diet. The model rats were divided into three groups as follows: finofibrate treatment group(finofibrate 80 mg?kg -1 po, once a day),simvastatin treatment group (simvastatin 4 mg?kg -1 po, once a day)and control group without either above-mentioned treatment. Experimental rats were treated for four weeks and then sacrificed for blood sampling. Serum free fatty acids were analyzed by gas chromatography. RESULTS Fenofibrate significantly ameliorated the decrease in polyunsaturated fatty acids induced by ethanol [oleic acid:(38.212?7.788) ?g?L -1 vs (31.620?6.142) ?g?L -1,linoleic acid:(37.269?8.065) ?g?L -1 vs (30.254?9.063) ?g?L -1,arachidonic acid:(11.646?2.601) ?g?L -1 vs (9.012?1.236) ?g?L -1] accompanied by the improvement of the fat infiltration of the liver, but demonstrated no effect on the increase in serum saturated fatty acids by ethanol. In the contrast, simvastatin can aggravate the decrease in polyunsatrurated fatty acids and significantly increase the levels of satrurated fatty acids in serum induced by ethanol along with the pathological aggravation of alcoholic fatty liver. CONCLUSION The results of present study revealed that fenofibrate and simvastatin exerted different effect on the serum free fatty acids of alcoholic fatty liver. Polyunsatrurated fatty acids in the serum play an important role in the pathogenesis and treatment response of alcoholic fatty liver.
9.Methodological research of endotracheal tube positioning confirmed by bedside ultrasound
Fenglin DONG ; Qingmin FAN ; Yan ZHENG ; Ajun WANG ; Lili ZHU ; Wei LEI
Chinese Journal of Ultrasonography 2016;25(4):309-312
Objective To evaluate the method of ultrasonic imaging to confirm endotracheal tube location in adult patients.Methods A certified sonographer identified the location of the trachea tube with ultrasound machine and then put it to optimum place.Correct trachea tube location confirmed by fiberoptic bronchoscopy (FOB) was used to evaluate the accuracy of ultrasonography for detecting endotracheal tube location.Results Several relevant structures,including anterior wall of trachea,the edge of balloon and the superior edge of the aorta (DGA) would be successfully visualized by sonographic examination.Among 48patients underwent ultrasound-guided tracheal intubation,there were 44 successful cases.2 intubatedmalposition cases and 2 failed-to-guide cases.The locating accuracy rate was 91.7%.Conclusions Ultrasound examination can identify the position of trachea tube in adult patients accurately,and it is a noninvasive,convenient and radiation free method for patients undergoing airway management.
10.Quality Surveillance of Sputum Specimen
Xiaoping LIU ; Yan JIANG ; Fan YANG ; Anping XU ; Jing DONG ; Mantian ZHANG
Chinese Journal of Nosocomiology 2009;0(16):-
OBJECTIVE To survey the qualification rate of sputum specimen.METHODS The specimen collecting and delivering time(morning,after deep coughing and gargling or not) was investigated.Aerobic bacteria isolated rate was evaluated.RESULTS The mean transportation time in positive aerobic bacteria isolated specimens and negative ones was 75 min and 124 min,respectively.Aerobic bacteria isolated rate was higher in sputum specimen that were microscopically screened for greater than 25 neutrophils,than in sputum specimen that were less then 25 neutrophils and greater than 10 BSE(buccal squamous epithelial) cells per 100? field.CONCLUSIONS Lower respiratory tract specimens should be delivered to the laboratory within 1 hour.Sputum specimen should be collected in the morning and after deep coughing and gargling.Microscopic examination should be mandatory in sputum microbiology,both for specimen evaluation and as a guide to what to look for in culture.